scholarly journals Development and Evaluation of a Duo Chikungunya Virus Real-Time RT-PCR Assay Targeting Two Regions within the Genome

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 755 ◽  
Author(s):  
Laurence Thirion ◽  
Laura Pezzi ◽  
Iban Corcostegui ◽  
Audrey Dubot-Pérès ◽  
Alessandra Falchi ◽  
...  
Keyword(s):  
Real Time ◽  
Chikungunya Virus ◽  
Point Mutations ◽  
In Silico Analysis ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Genetic Lineages ◽  
Sensitivity Specificity ◽  
Dual Target ◽  
Major Sequence

Chikungunya virus (CHIKV) re-emerged as a globalized health threat fifteen years ago. There are dozens of RT-PCR assays published. An inventory of the latter was made, and after in silico analysis, two assays were selected for their ability to detect strains belonging to the five CHIKV genetic lineages. They were combined in order to provide a robust assay not affected by genetic point mutations and the resulting Duo CHIKV real-time RT-PCR assay was compared to the two parental single-plex tests against five strains belonging to the five genetic lineages. The Duo CHIKV assay performed equally, or better, in terms of sensitivity, specificity, linearity and signal intensity. Dual-target assays are better suited for viruses having the propensity to evolve into new variants via point mutations or major sequence deletions/insertions. Here, we demonstrated that combining two single systems into a dual-target assay did not impair sensitivity and specificity, and proved a potent diagnostic tool to face a potential emergence of CHIKV variants by newly evolving mutations.

Detection and Quantification of Chikungunya Virus by Real-Time RT-PCR Assay

2016 ◽  
pp. 105-117 ◽  
Author(s):  
Seok Mui Wang ◽  
Ummul Haninah Ali ◽  
Shamala Devi Sekaran ◽  
Ravindran Thayan
Keyword(s):  
Real Time ◽  
Chikungunya Virus ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Detection And Quantification

scholarly journals Development and evaluation of one-step multiplex real-time RT-PCR assay for simultaneous detection of Zika virus and Chikungunya virus

2017 ◽  
Vol 90 (3) ◽  
pp. 389-396 ◽  
Author(s):  
Si-Qing Liu ◽  
Xiao Li ◽  
Cheng-Lin Deng ◽  
Zhi-Ming Yuan ◽  
Bo Zhang
Keyword(s):  
Real Time ◽  
Zika Virus ◽  
Chikungunya Virus ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
One Step

scholarly journals In-house standardization and validation of a multiplex RT-PCR assay for the detection of 13 respiratory viruses

Nova ◽  
2016 ◽  
Vol 14 (26) ◽  
pp. 9
Author(s):  
Hernán Vargas ◽  
Ángela Diaz ◽  
Yamile Celis ◽  
Liliana Díaz ◽  
Sandra Gómez ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
Real Time ◽  
Real Time Pcr ◽  
Respiratory Viruses ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Lower Efficiency ◽  
Single Target ◽  
Syncytial Virus ◽  
Sensitivity Specificity

<p>Background. Multiplex real time PCR is increasingly used to diagnose respiratory viruses and has shown to be superior to traditional methods, such as culture and antigen detection. Objective. Standardization and validation of a multiplex real-time PCR assay for the detection of 13 respiratory viruses. Methods. The assay was validated using RNA control targets and comparing results to single-target PCR’s. Results. Using RNA controls the multiplex format was found to be as sensitive and specific as the single-target PCRs, and no competition was observed between targets. The efficiencies for most of the reactions were approximately 90%, but a lower efficiency was found for Parainfluenza 2 with a rate of amplification in each cycle of 86.63%. On the other hand, a higher efficiency was observed in respiratory syncytial virus A and respiratory syncytial virus B ((93.07% each). Conclusion: This multiplex RT-PCR format shows an adequate efficiency, demonstrating an excellent sensitivity, specificity and repeatability for all the studied respiratory viruses.</p>

Development and evaluation of SYBR Green I-based one-step real-time RT-PCR assay for detection and quantification of Chikungunya virus

2007 ◽  
Vol 39 (3) ◽  
pp. 188-193 ◽  
Author(s):  
S.R. Santhosh ◽  
M.M. Parida ◽  
P.K. Dash ◽  
A. Pateriya ◽  
B. Pattnaik ◽  
...  
Keyword(s):  
Real Time ◽  
Chikungunya Virus ◽  
Sybr Green ◽  
Sybr Green I ◽  
Rt Pcr ◽  
Pcr Assay ◽  
One Step ◽  
Detection And Quantification

scholarly journals A35 The first laboratory confirmation of chikungunya outbreak in Ethiopia

2019 ◽  
Vol 5 (Supplement_1) ◽  
Author(s):  
Mesfin Mengesha Tsegaye ◽  
Aadamu Tayachew ◽  
Desalegn Belay ◽  
Abebe Alemu ◽  
Berhane Beyene
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Laboratory Investigation ◽  
Emergency Preparedness ◽  
Zika Virus ◽  
Chikungunya Virus ◽  
Febrile Illness ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Serum Samples

Abstract Chikungunya is a viral disease (genus Alphavirus) which is transmitted to humans by infected mosquitoes—including Aedes aegypti and A. albopictus. An outbreak of febrile illness, suspected to have been caused by chikungunya, was reported in June 2016 from Dolloado district, Suuf Kebele, in the Somalia regional state of Ethiopia that borders the Mandera county of Kenya where a confirmed chikungunya outbreak was ongoing. Laboratory investigation was carried out to confirm if the outbreak in Ethiopia was caused by Chikungunya virus. Ten serum samples were collected from suspected patients visiting a health center in Suuf Kebel, who were then sent to the Nation laboratory in Ethiopian Public Health Institute. RNA was extracted from the serum samples using QIAgene RNA Mini kit, and PCR detection of dengue, chikungunya, and Zika virus nucleic acid was done using Trioplex Real-time RT-PCR Assay following the protocol from the Center for Disease Control (CDC). The Trioplex Real-time RT-PCR assay, for detection and differentiation of RNA from dengue, Chikungunya and Zika, was provided by CDC as part of the zika emergency preparedness effort. Of the nine samples tested, eight (88.88%) were found to be positive for chikungunya virus nucleic acid but negative for dengue and Zika virus nucleic acids. The median age of the affected sampled patients was 40 years, and males appear to be more affected (66.6% of sampled patients). The laboratory investigation confirmed that the outbreak was caused by chikungunya virus. Even though further molecular characterization of the positive isolates will provide more information as to the circulating genotypes and elucidate the origin of the outbreak virus, it is also possible to assume that the outbreak was an extension of the outbreak in neighboring countries in Kenya and, therefore, warrants that cross-border integration efforts to control chikungunya should be implemented by the concerned countries.

scholarly journals Development and application of a real-time RT-PCR assay to rapidly detect H2 subtype avian influenza A viruses

2021 ◽  
pp. 104063872199481
Author(s):  
Yixin Xiao ◽  
Fan Yang ◽  
Fumin Liu ◽  
Hangping Yao ◽  
Nanping Wu ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Influenza A ◽  
Minor Groove Binder ◽  
Rt Pcr ◽  
Influenza A Viruses ◽  
Pcr Assay ◽  
Infectious Dose ◽  
The Matrix ◽  
Taqman Minor Groove Binder

The H2 subtypes of avian influenza A viruses (avian IAVs) have been circulating in poultry, and they have the potential to infect humans. Therefore, establishing a method to quickly detect this subtype is pivotal. We developed a TaqMan minor groove binder real-time RT-PCR assay that involved probes and primers based on conserved sequences of the matrix and hemagglutinin genes. The detection limit of this assay was as low as one 50% egg infectious dose (EID50)/mL per reaction. This assay is specific, sensitive, and rapid for detecting avian IAV H2 subtypes.

A one-step multiplex real-time RT-PCR assay for rapid and simultaneous detection of human norovirus genogroup I, II and IV

2013 ◽  
Vol 189 (2) ◽  
pp. 277-282 ◽  
Author(s):  
Yong Yan ◽  
Heng-hui Wang ◽  
Lei Gao ◽  
Ji-mei Ji ◽  
Zhi-jie Ge ◽  
...  
Keyword(s):  
Real Time ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Human Norovirus ◽  
One Step

Multicenter evaluation of the ENTEROVIRUS R-gene™ real-time RT-PCR assay for the detection of enteroviruses in clinical specimens

2010 ◽  
Vol 47 (1) ◽  
pp. 54-59 ◽  
Author(s):  
Sylvie Pillet ◽  
Geneviève Billaud ◽  
Shabir Omar ◽  
Bruno Lina ◽  
Bruno Pozzetto ◽  
...  
Keyword(s):  
Real Time ◽  
R Gene ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Clinical Specimens
Sign in / Sign up

Export Citation Format

Share Document